| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note | 
|---|---|---|---|---|---|---|---|---|
| 2 |  Lake Highland Prep | 
		A | Alicea | DeMartino | Parekh | Vidrine | NT | |
| 3 |  Tohopekaliga | 
		A | Cruz | Francois | Richey | Medina | 4:36.32 | 4:36.32 St. Cloud Invitational | 
| 4 |  Jones | 
		A | Bryant | Dorns | Hall | Yarn | 4:22.03 | 4:22.03 Bishop Moore - Larry Kelly Invite 2024 %28Regional Qualifier%29 | 
| 5 |  Oviedo | 
		A | Atwell | Wahl | Schnoor | Price | 4:13.95 | FHSAA 4A District 3 | 
| 6 |  Mainland | 
		A | Rivers | Nixon | Polk | Williams | 4:17.00 | |
| 7 |  Winter Park | 
		A | Arriola | Leon | Guitton | Denney | 4:31.97 | 4:31.97 Charlie Harris Relays | 
| 8 |  Trinity Prep | 
		A | Burgos | Patino | Macau | Hennebery | DNS |